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Parasite (Paris, France) 2022Commercial multiplex PCR assay panels were developed to overcome the limitations of microscopic examination for parasitological diagnosis on stool samples. However,...
Selecting a multiplex PCR panel for accurate molecular diagnosis of intestinal protists: a comparative study of Allplex (Seegene), G-DiaParaTrio (Diagenode), and RIDAGENE (R-Biopharm) assays and microscopic examination.
Commercial multiplex PCR assay panels were developed to overcome the limitations of microscopic examination for parasitological diagnosis on stool samples. However, given the increased supply of this diagnostic approach, these assays must be evaluated to position them in a diagnostic algorithm. Analytical performances of the multiplex PCR assay G-DiaParaTrio, Allplex GI parasite and RIDAGENE parasitic stool panel for detecting Blastocystis sp., Entamoeba histolytica, Giardia duodenalis, Cryptosporidium spp., Dientamoeba fragilis, and Cyclospora cayetanensis, were assessed through a retrospective comparative study on 184 stool samples initially sent for parasitological investigation. The composite reference method for parasitological diagnosis was microscopic observation and Entamoeba histolytica-specific adhesion detection when necessary. Multiplex PCR assays were performed on extracted DNA from each stool, following the manufacturer's recommendations. Discrepant results with the composite reference method were investigated with species-specific PCR to approach a final parasitological diagnosis. Overall sensitivity/specificity for the multiplex PCR assays was 93.2%/100% for G-DiaParaTrio, 96.5%/98.3% for Allplex GI parasite and 89.6%/98.3% for RIDAGENE, whereas the composite reference method presented an overall sensitivity/specificity of 59.6%/99.8%. These results confirmed the added diagnostic value of the multiplex PCR approach for gastrointestinal protists. Nevertheless, the PCR procedure and the analytical performance for each protist of interest, variable depending on the multiplex PCR assay, must be considered when implementing a PCR-based diagnostic approach.
Topics: Animals; Cryptosporidiosis; Cryptosporidium; Entamoeba histolytica; Feces; Giardia lamblia; Multiplex Polymerase Chain Reaction; Retrospective Studies; Scrapie; Sensitivity and Specificity; Sheep
PubMed: 35138245
DOI: 10.1051/parasite/2022003 -
Journal of Travel Medicine Jul 2022Persistent abdominal symptoms (PAS) are the leading cause of post-travel morbidity although there is a paucity of evidence concerning the aetiology of this condition....
BACKGROUND
Persistent abdominal symptoms (PAS) are the leading cause of post-travel morbidity although there is a paucity of evidence concerning the aetiology of this condition. Recently molecular methods for protozoa detection in stool have been introduced. Herein, we describe the clinical aspects and the prevalence of gastrointestinal protozoa in returning travellers with PAS.
METHODS
From 2017 to 2019, clinical information and stool specimens from returning travellers with PAS were analysed for the presence of parasites using the Allplex-GI-Parasite-assay. Stool findings from symptomatic patients without a travel history were used as a comparator.
RESULTS
During the 2-year study, 203 stool specimens from returning travellers were analysed. The median duration of symptoms before seeking care was 6 months, the most common symptoms were fatigue (79.2%), abdominal pain (75.7%) and loose stool (70.8%).Most of travellers had returned from Asia (57.6%), mainly from the Indian-subcontinent and only 52.6% were backpackers. Altogether, 36.9% samples were positive for protozoa, with Blastocystis hominis being the most common (26.6%) in samples, followed by Dientamoeba fragilis (18.7%), Giardia lamblia (3.0%) and Cryptosporidium spp (0.5%). The former two were dominant in all regions. In all cases but one, G. lamblia was acquired, but one were acquired in the Indian subcontinent (odds ratios 16.9; 95% confidence intervals: 1.9-148.3). Entamoeba histolytica was not detected. The demographic characterization of the 1359 non-travellers was comparable with the travellers. Among them D. fragilis was the most common followed by B. hominis, which was significantly less frequent compared among the travellers (16.7% vs 26.6%, P < 0.001). Average Cycle threshold values for each stool parasites were comparable between the two groups.
CONCLUSION
Among returning travellers with PAS, more than one-third were positive for gastrointestinal protozoa. A low rate of giardia was found and no E. histolytica while B. hominis followed by D. fragilis were the dominant findings. Further studies are required to better understand the role of these protozoa in PAS.
Topics: Cryptosporidiosis; Cryptosporidium; Entamoeba histolytica; Feces; Giardia lamblia; Giardiasis; Humans
PubMed: 35134178
DOI: 10.1093/jtm/taac011 -
European Journal of Gastroenterology &... Feb 2022
Topics: Blastocystis; Dientamoeba; Feces; Freezing; Humans; Irritable Bowel Syndrome
PubMed: 34845142
DOI: 10.1097/MEG.0000000000002327 -
International Journal of Obesity (2005) Jan 2022Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic...
BACKGROUND
Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic parasites in this microenvironment is still largely unknown. Their presence within the gut ecosystem in obese subjects warrants further study, as well as their influence on the host metabolism and comorbidities.
METHODS
Herein, a single center, cross-sectional study of 104 obese individuals was performed to assess the presence of six intestinal unicellular parasites in stool using a commercially available kit, and to evaluate its relationship with the presence of abdominal symptoms, metabolic comorbidities, variations in body composition and nutritional deficiencies.
RESULTS
The overall parasitic colonization rate was 51%, with Blastocystis sp., identified as the most frequent (44.2%), followed by Dientamoeba fragilis (11.5%) and Giardia intestinalis (8.7%), and significantly related to the consumption of ecological fruits and vegetables. Contrary to what previous studies pointed out, colonization with parasites species was significantly associated with fewer abdominal symptoms and depositions per day. The presence of parasites did not correlate with any nutritional deficiencies nor differences in body composition, while it did with significant lower HOMA-IR levels and a lower trend towards metabolic syndrome.
CONCLUSION
Obese subjects frequently harbor unicellular enteric parasites, apparently without clinical nor nutritional harm. This evidence suggests that carrying these microorganisms, from an endocrinological perspective, has a beneficial effect, especially on insulin resistance and possibly on the development of related comorbidities.
Topics: Adult; Animals; Blastocystis; Cross-Sectional Studies; Female; Gastrointestinal Tract; Humans; Male; Obesity, Morbid; Parasites; Spain
PubMed: 34650200
DOI: 10.1038/s41366-021-00980-6 -
Case Reports in Gastroenterology 2021Budd-Chiari syndrome (BCS) is an uncommon illness that is characterized by obstruction of hepatic venous outflow. Patients typically present with nausea, vomiting, and...
Budd-Chiari syndrome (BCS) is an uncommon illness that is characterized by obstruction of hepatic venous outflow. Patients typically present with nausea, vomiting, and abdominal pain, which can further progress into signs associated with liver failure, including jaundice, encephalopathy, and coagulopathy. The most common causes of BCS include pathologies that induce portal vein thrombosis, such as myeloproliferative disorders, malignancy, and acquired hypercoagulable states. In this case report, a patient who presented with abdominal pain and distention is diagnosed with BCS caused by an unusual etiology. He was found to have significant eosinophilia, prompting additional evaluation for parasitic infections. Using stool diagnostics/studies, he was found to have trophozoites. The patient was treated with enoxaparin, warfarin, and metronidazole with a resolution of his symptoms. This case outlines a novel cause of BCS as well as the proposed mechanism of induction of BCS.
PubMed: 34616241
DOI: 10.1159/000516210 -
PLoS Neglected Tropical Diseases Oct 2021The clinical significance of Blastocystis sp. and Dientamoeba fragilis in patients with gastrointestinal symptoms is a controversial issue. Since the pathogenicity of...
Investigation of neglected protists Blastocystis sp. and Dientamoeba fragilis in immunocompetent and immunodeficient diarrheal patients using both conventional and molecular methods.
INTRODUCTION
The clinical significance of Blastocystis sp. and Dientamoeba fragilis in patients with gastrointestinal symptoms is a controversial issue. Since the pathogenicity of these protists has not been fully elucidated, testing for these organisms is not routinely pursued by most laboratories and clinicians. Thus, the prevalence of these organisms and the subtypes of Blastocystis sp. in human patients in Turkey are not well characterized. This study aimed to determine the prevalence of Blastocystis sp. and D. fragilis in the diarrheic stool samples of immunodeficient and immunocompetent patients using conventional and molecular methods and to identify Blastocystis sp. subtypes using next generation sequencing.
MATERIAL AND METHODS
Individual stool specimens were collected from 245 immunodeficient and 193 immunocompetent diarrheic patients between March 2017 and December 2019 at the Gazi University Training and Research Hospital in Ankara, Turkey. Samples were screened for Blastocystis sp. and D. fragilis by conventional and molecular methods. Molecular detection of both protists was achieved by separate qPCRs targeting a partial fragment of the SSU rRNA gene. Next generation sequencing was used to identify Blastocystis sp. subtypes.
RESULTS
The prevalence of Blastocystis sp. and D. fragilis was 16.7% and 11.9%, respectively as measured by qPCR. The prevalence of Blastocystis sp. and D. fragilis was lower in immunodeficient patients (12.7% and 10.6%, respectively) compared to immunocompetent patients (21.8% and 13.5%, respectively). Five Blastocystis sp. subtypes were identified and the following subtype distribution was observed: ST3 54.4% (n = 37), ST2 16.2% (n = 11), ST1 4.4% (n = 3), ST6 2.9% (n = 2), ST4 1.5% (n = 1), ST2/ST3 11.8% (n = 8) and ST1/ST3 8.8% (n = 6). There was no statistically significant difference in the distribution of Blastocystis sp. subtypes between immunocompetent and immunodeficient patients.
CONCLUSION AND RECOMMENDATION
Our findings demonstrated that Blastocystis sp. and D. fragilis are commonly present in immunocompetent and immunodeficient patients with diarrhea. This study is the first to use next generation sequencing to address the presence of Blastocystis sp. mixed subtypes and intra-subtype variability in clinical samples in Turkey.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blastocystis; Diarrhea; Dientamoeba; Feces; Female; High-Throughput Nucleotide Sequencing; Humans; Immunocompetence; Male; Middle Aged; Primary Immunodeficiency Diseases; Turkey; Young Adult
PubMed: 34613993
DOI: 10.1371/journal.pntd.0009779 -
PLoS Neglected Tropical Diseases Sep 2021Childhood diarrhoea, a major cause of morbidity and mortality in low-income regions, remains scarcely studied in many countries, such as Guinea-Bissau. Stool sample...
BACKGROUND
Childhood diarrhoea, a major cause of morbidity and mortality in low-income regions, remains scarcely studied in many countries, such as Guinea-Bissau. Stool sample drying enables later qPCR analyses of pathogens without concern about electricity shortages.
METHODS
Dried stool samples of children under five years treated at the Bandim Health Centre in Bissau, Guinea-Bissau were screened by qPCR for nine enteric bacteria, five viruses, and four parasites. The findings of children having and not having diarrhoea were compared in age groups 0-11 and 12-59 months.
RESULTS
Of the 429 children- 228 with and 201 without diarrhoea- 96.9% and 93.5% had bacterial, 62.7% and 44.3% viral, and 52.6% and 48.3% parasitic pathogen findings, respectively. Enteroaggregarive Escherichia coli (EAEC; 60.5% versus 66.7%), enteropathogenic E. coli (EPEC; 61.4% versus 62.7%), Campylobacter (53.2% versus 51.8%), and enterotoxigenic E. coli (ETEC; 54.4% versus 44.3%) were the most common bacterial pathogens. Diarrhoea was associated with enteroinvasive E. coli (EIEC)/Shigella (63.3%), ETEC (54.4%), astrovirus (75.0%), norovirus GII (72.6%) and Cryptosporidium (71.2%). The only pathogen associated with severe diarrhoea was EIEC/Shigella (p<0.001). EAEC was found more frequent among the infants, and EIEC/Shigella, Giardia duodenalis and Dientamoeba fragilis among the older children.
CONCLUSIONS
Stool pathogens proved common among all the children regardless of them having diarrhoea or not.
Topics: Bacteria; Bacterial Infections; Child, Preschool; Diarrhea; Feces; Female; Guinea-Bissau; Humans; Infant; Male; Virus Diseases; Viruses
PubMed: 34587158
DOI: 10.1371/journal.pntd.0009709 -
Journal of Clinical Microbiology Nov 2021Despite the global use of rotavirus vaccines, vaccine breakthrough cases remain a pediatric health problem. In this study, we investigated suspected rotavirus vaccine...
Despite the global use of rotavirus vaccines, vaccine breakthrough cases remain a pediatric health problem. In this study, we investigated suspected rotavirus vaccine breakthrough cases using next-generation sequencing (NGS)-based viral metagenomics ( = 102) and a panel of semiquantitative reverse transcription-PCR (RT-qPCR) ( = 92) targeting known enteric pathogens. Overall, we identified coinfections in 80% of the cases. Enteropathogens such as adenovirus (32%), enterovirus (15%), diarrheagenic Escherichia coli (1 to 14%), astrovirus (10%), spp. (10%), parechovirus (9%), norovirus (9%), Clostridioides (formerly ) difficile (9%), Dientamoeba fragilis (9%), sapovirus (8%), Campylobacter jejuni (4%), and Giardia lamblia (4%) were detected. Except for a few reassortant rotavirus strains, unusual genotypes or genotype combinations were not present. However, in addition to well-known enteric viruses, divergent variants of enteroviruses and nonclassic astroviruses were identified using NGS. We estimated that in 31.5% of the patients, rotavirus was likely not the cause of gastroenteritis, and in 14.1% of the patients, it contributed together with another pathogen(s) to disease. The remaining 54.4% of the patients likely had a true vaccine breakthrough infection. The high prevalence of alternative enteropathogens in the suspected rotavirus vaccine breakthrough cases suggests that gastroenteritis is often the result of a coinfection and that rotavirus vaccine effectiveness might be underestimated in clinical and epidemiological studies.
Topics: Child; Feces; Humans; Prevalence; Real-Time Polymerase Chain Reaction; Rotavirus Vaccines
PubMed: 34586890
DOI: 10.1128/JCM.01236-21 -
Tropical Medicine and Infectious Disease Sep 2021While the prevalence of intestinal parasitic infections (IPI) has been most commonly studied in African and Asian populations, less is known about the prevalence rates... (Review)
Review
While the prevalence of intestinal parasitic infections (IPI) has been most commonly studied in African and Asian populations, less is known about the prevalence rates of IPI in European children, as well as the potential risk factors that favor the spread of parasites. We aimed to review published evidence on the prevalence rates of IPI in children residing in Europe, and to quantitatively synthesize the results of published studies. We searched Medline from 1 January 2015 to 1 April 2021 to address the most recently published prevalence patterns of IPI in European children. Random-effects meta-analyses were performed by type of IPI infection, age group and sex, depending on data availability. Of the 967 potentially relevant articles, eight eligible cross-sectional studies were included in this analysis, yielding a sample of 3376 children (0-19 years). The overall prevalence rate was 5.9% for any IPI in children residing in European countries. was the most commonly detected parasite yielding a prevalence rate of 10.7%. Other parasites included and . Studies focusing on specific types of parasites showed prevalence rates ranging from 1.3% for to 68.3% for . Despite the scarce literature, the present review showed relatively low prevalence rates of IPI in Europe. Future studies accounting for proper diagnostic methods used for the detection of parasites and including information on potential sociodemographic factors, such as travelling history and history of immigration, are needed to guide clinicians about which children to test, as well as when and how to test children for IPI.
PubMed: 34564544
DOI: 10.3390/tropicalmed6030160 -
Journal of Pediatric Gastroenterology... Dec 2021This survey was undertaken to obtain insight in the attitude of Dutch physicians towards pathogenicity, diagnostic- and therapeutic approach towards Dientamoeba fragilis...
This survey was undertaken to obtain insight in the attitude of Dutch physicians towards pathogenicity, diagnostic- and therapeutic approach towards Dientamoeba fragilis in children. Physicians were invited by e-mail for a questionnaire. A total of 211 of 450 physicians (46.9%) completed the questionnaire, including 67 general practitioners (GPs) and 144 pediatricians. Of all respondents, 175 of 211 (82.9%) considered D fragilis a "potential pathogen", when other causes of gastro-intestinal complaints are ruled out. Only 16 of 211 (7.6%) performed diagnostic tests regularly. Diagnostic tests were performed by 162 of 211 (77%) of respondents in children with diarrhea and abdominal pain in consideration of duration of symptoms. Fecal polymerase chain reaction (PCR) was diagnostic modality of preference. Eighty-nine of 142 (62.7%) prescribed metronidazole as antibiotic of first choice. This study shows heterogeneity in clinical practice amongst Dutch physicians regarding diagnostic- and therapeutic approach of D fragilis in children. Different attitude towards pathogenicity and inconsistent guidelines could be causative factors.
Topics: Child; Dientamoeba; Dientamoebiasis; Feces; General Practitioners; Humans; Netherlands; Pediatricians; Surveys and Questionnaires
PubMed: 34520404
DOI: 10.1097/MPG.0000000000003297